USPTO Patents Application 09993647

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Artic l e T00I3

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Alteration of lacto-series glycolipid glycosyltransferase activities
in human colonic adenocarcinoma DLD-1 cells after culture in
N,N-dimethylformamide-containing medium.

J Cell Biochem.
2312)

1990: 44(2):93-105 (ISSN: 0730-

Holmes EH; Greene TG

Pacific Northwest Research Foundation, Seattle, Washington
98122.

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Human colonic adenocarcinoma DLD-1 cells were grown under
conditions which induce characteristics of differentiated cells using
medium containing 0.8% N,N-dimethylformamide in order to study
alterations in glycosphingolipid glycosyltransferase activities
during this process. Analysis of biosynthetic reactions involved in
lacto-series antigen synthesis revealed no changes in the specific
activities of either beta 1 — 4galactosyltransferase or alpha 1 —
3/4fucosyltransferase with N,N-dimethylformamide treatment.
However, a dramatic decrease of from 14- to 20-fold in the beta 1-
— 3N-acetylglucosaminyltransferase activity was observed in the
treated cells. This enzyme catalyzes the rate-limiting step in lacto-
series core chain synthesis. This is consistent with the pattern of
regulation of lacto-series antigen expression found to occur during

oncogenesis in human colonic mucosa (Holmes EH, Hakomori S, Ostrander GK: J Biol Chem 262:15649,
1987). Total glycolipids from untreated and N,N-dimethylformamide-treated cells were isolated and
subjected to TLC immunostain analysis and solid phase radioimmunoassay with a series of monoclonal
antibodies specific for lacto-series-based carbohydrate antigens. A decrease of about 2-fold or less in the
quantity of lacto-series antigens was observed as a consequence of N,N-dimethylformamide treatment in
both neutral glycolipid and ganglioside fractions. The results suggest that only very low levels of beta 1 —
3N-acetylglucosaminyltransferase activity are required for the steady state expression of significant levels
of lacto-series based glycolipids and that modulation of its activity levels by N,N-dimethylformamide
treatment in DLD-1 cells represents a convenient in vitro system for studying aspects of regulation of
lacto-series antigen expression.

^ Subject Headings

PreMedline Identifier: 2250046

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